Method for preventing periodontitis

ABSTRACT

The use of stabilized chlorine dioxide or chlorine dioxide in aqueous solution as a composition to reduce or prevent plaque formation and treatment of oral diseases ecologically plaque dependent, such as gingivitis and periodontitis, is disclosed. Preferred concentrations are in the range of 0.005% to 0.2% and the chlorine dioxide may be in the form of a rinse, a wash, a soak or a dentrifice.

REFERENCE TO DETAILED APPLICATION

This application is a divisional application of a copending applicationentitled "IMPROVED METHOD AND COMPOSITION FOR PREVENTION OF PLAQUEFORMATION AND PLAQUE DEPENDENT DISEASES," Ser. No. 088,609, filed Aug.24, 1987, now U.S. Pat. No. 4,818,519, which is a continuation in partapplication of a copending application entitled "METHOD AND COMPOSITIONFOR PREVENTION OF PLAQUE FORMATION AND PLAQUE DEPENDENT DISEASES", Ser.No. 024,329, filed Mar. 10, 1987, now U.S. Pat. No. 4,837,009 which is acontinuation application of copending applications entitled "METHOD ANDCOMPOSITION FOR PREVENTION AND TREATMENT OF ORAL DISEASE", Ser. No.947,079, filed Dec. 29, 1986, now U.S. Pat. No. 4,689,215, issued Aug.25, 1987, and Ser. No. 017,241, filed Dec. 29, 1986, now U.S. Pat. No.4,696,811, issued Sept. 2, 1987, which applications are continuationapplications of copending application "METHOD AND COMPOSITION FORPREVENTION AND TREATMENT OF ORAL DISEASE", Ser. No. 846,342, filed Mar.31, 1986 (now abandoned), which is a continuation application ofcopending application entitled "METHOD AND COMPOSITION FOR PREVENTIONAND TREATMENT OF ORAL DISEASE", Ser. No. 636,027, filed July 30, 1984(now abandoned).

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention is directed to a method and composition forprevention and treatment of oral disease and more particularly, to theuse of a chlorine dioxide in aqueous solution for preventing formationof plaque and treatment of plaque dependent diseases.

2. Description of the Prior Art

The volatile sulfur compounds, hydrogen sulfide (H₂ S) methylmercaptan(CH₃ SH) and di-methylmercaptan (CH³)₂ S are recognized in the currentdental literature as being the major contributors to oral malodor.Numerous researchers using organoleptic, chemical, amperometric, massspectrometric, or gas liquid chromatographic methods have demonstratedthat these volatile sulfur compounds are present in the head space andvapor of putrefied saliva and in individual samples of mouth air. Inmost persons, hydrogen sulfide and methylmercaptan constitute over 90%of the total volatile sulfur content identified in mouth air.

These malodorous volatile sulfur compounds are generated primarilythrough the putreficative action of oral microorganisms on sulfurcontaining amino acids, peptones or proteins found in the mouth. Thesesubstrates are readily available in saliva and dental plaque or may bederived form proteinaceous food particles trapped between the teeth, inthe gingival crevice or adhering to the mucous membranes and theirregular surface of the tongue as well as exfoliated oral epithelium,food debris and the like. Current studies have indicated that mouth odornot only comes from the posterior dorsal surface of the tongue but alsofrom periodontal pockets. People with periodontal involvement have anattendant increase in oral malodor from disintegrated epithelial cells.

Starting with a clean tooth surface, plaque formation and resultingecology occurs in the following steps:

1. Deposition of a coating of glycoproteins from salivary and other oralmucous gland secretions. This is referred to as acquired pellicle.

2. Fastening and colonization of streptococcus organisms to the acquiredpellicle, primarily by streptococcus sanguis and streptococcus mutans.

3. Conversion of sucrose to glucans (dextran) and fructans by thebacterial enzyme glucosyltransferases. In this plaque mass are imbeddeddead cells, cell debris and food debris. High molecular weight polymersof glucose and other sugars, altered salivary glycoproteins, proteasesand various chemotactic and inflammatory inducing substances have beendetected and partially characterized.

4. Other organisms, primarily gram positive aerobes, become residents inthe plaque mass and use the glucans and fructans for nutrition. Theseare primarily oxygen using organisms and the oxygen source is from thesaliva that bathes the plaque mass.

5. With time and the functioning of this ecological system, the oxygenuse by the superficial bacteria deprive the lower layers of the plaquematrix of a supply of oxygen. An opportunity for non-oxygen usingbacteria (facultative anaerobes) to become established is provided.

6. If left undisturbed, the ecological system now established is selfperpetuating. That is, the streptococcus bacteria continue to produceglucans and fructans. Other bacteria produce toxins that kill cells ofthe host and the dead cells become other essential nutrients. Thesuperficial bacteria deprive the deeper layers of the plaque mass ofoxygen and keep the ecological system going. Thus, both aerobic andanaerobic organisms survive int he plaque mass.

7. The established ecological system attendant the plaque mass producestoxins from the aerobic bacteria that cause gingivitis and toxins fromthe anaerobic bacteria that cause periodontitis.

Various substances have been tested for their ability to disrupt plaqueor prevent its formation and to treat mouth odor such as antibiotics,chlorhexdines, oxine, and alexidine.

The prior art compositions that have been used and tested, have foundsome acceptance but are generally inefective in periodontitis,gingivitis, plaque accumulation and mouth malodor. Accordingly, thereexists a clear need for composition which will effectively inhibit theinitial pellicle which precedes plaque formation and inhibit or controlthe formation of bacterial plaque and suppress organisms such as but notlimited to (1) Streptococcus mutans, which is implicated as the majorcause of human dental decay; (2) Black Pigmented Bacteriodes, anActinobacillus actinomycetumcomitans which is implicated in humanperiodontitis; and (3) will reduce odor intensity in the mouth throughthe control of hydrogen sulfide and methylmercaptan.

SUMMARY OF THE PRESENT INVENTION

Broadly, the present invention contemplates the use of stabilizedchlorine dioxide in aqueous solution for the treatment of the mouth as adeodorizing agent, anti-plaque agent, bactericide for treatment ofgingivitis and periodontitis and as a bactericidal fungicidal andviralcidal agent in other related applications. In the present inventiona composition containing stabilized chlorine dioxide may be used fortreatment of the mouth either in a solution, for example, as a mouthwashor in a dentifrice generally in concentrations of below approximately0.2% for the control of odorgenic microorganisms, bacterial plaque,gingivitis and bacteria which cause these conditions. Similarly,chlorine dioxide is also effective as a cellular debridgement agentfollowing surgical procedures and sanitizer denture soak. The use ofchlorine dioxide and its effects on man has been clinically evaluated.The relative safety of oral ingestion of chlorine dioxide wasdemonstrated extensively in animals and latter in humans by Lubbers,Chauan, and Bianchine, Environmental Health Perspectives, Volume 46,Pages 57-62, 1982.

DESCRIPTION OF PREFERRED EMBODIMENTS

Chlorine dioxide, ClO₂, functions biochemically in many ways other thanas a germicide. These functions include: (1) oxidation of double bondsbetween two carbon atoms; (2) oxidation of unsaturated fatty acids(lipids) via double bonds between two carbon atoms; (3) acceleration ofhydrolysis of carboxalic anhydrides; (4) oxidation of aldehydes to thecorresponding carboxalic acids; (5) oxidation of alcohols; (6) oxidationof amines; (7) oxidation of phenols, phenolic derivatives andthiophenolic compounds; (8) moderate oxidation of hydroquinones; (9)oxidation of thiophenols; (10) Oxidation of amino acids, proteins andpolyamides, primarily by attacking sulphide bonds. These are cystine,methionone and tryrosine. Tryptophane also has been shown to bereactive. Keratin, (which makes up the cyto-skeletal structure inepithelial cells cytoplasm), and ClO₂ keratin sulfonic hydrosolubleacids; (11) carbohydrates are altered at the CHO and CH₂ OH radicals toproduce carboxylic functions; and (12) Nitrates and sulphides arereadily oxidized.

The chlorine dioxide described herein is of the type referred to asstabilized chlorine dioxide. U.S. Pat. No. 3,271,242 describes a form ofstabilized chlorine dioxide and a method of making it which isparticularly useful in carrying out the present invention. Furtherdiscussion of stabilized chlorine dioxide in a form contemplated by thepresent invention may be found in a treatise entitled Chlorine Dioxideby W. J. Masschelein and published by the Ann Arbor Science Publishers,Inc., copyright 1979 (note in particular pages 138-140). Variousembodiments of chlorine dioxide for various purposes are also reviewedin this treatise.

The first step in the formation of plaque on a clean tooth surface isthe formation of acquired pellicle. Studies by others have shown thefollowing to be part of the acquired pellicle formative process.Glycoproteins of salivary and other mucous gland origin are attached tothe hydroxyapatite crystals. (Roukima, P. A. and Nieuw Amerongen, A. V.,Sulphated Glycoproteins in Human Saliva; Saliva and Dental Caries, (Sp.Supp. Microbiology Abst.) 1979, pp 76. Embery, G., The role of anionicglyco-conjugates, particularly Sulphated Glycoproteins in relation tothe Oral Cavity, Saliva and Dental Caries, (Supp. Microbiol Abstr.),Information Retrieval 1978, pp 105-111). Sulphated glycoproteins have astrong affinity to the calcium anion (ibid., pp 105-108). Most majorsalivary secreted glycoproteins may be bound to certain ester sulphates(ibid). These sulphated glycoproteins have been related to bacterialagglutination or clumping (ibid., pp 108).

Clinical observations by the inventor have led to the discovery that theprocess of acquired pellicle can be inhibited bythe use by humans ofstabilized chlorine dioxide as a rinse. Through such observations is hasbeen learned that the chlorine dioxide reacts with the sulphatedglycoproteins to inhibit pellicle formation. This process resultsprimarily from, but is not limited to, oxidation of the sulphide bonds.Since acquired pellicle is the first step in plaque formation, thisinitial inhibition alters the sequence of events to follow. The secondstep, bacterial adhesion and subsequent steps are consequently retarded.No disulphate enzymes capable of cleaning the sulphate moieties ofglycoproteins are known.

Bacterial agglutinigation includes the conversion of sucrose to glucansand fructans by enzymes known as glycosyltransferases. These enzymes areof bacterial origin. The plaque mass becomes a complex extra cellular(of microorganisms) matrix containing sulphated glucosamineglycans,proteoglycans, glycoproteins, sugar, proteins and lipids which aid inthe process of bacterial agglutination (Schluger, S., Yuodelis, R. andPage, R., Periodontal Disease, Chapter 6, pp. 135-166, 1977, Lea &Febiger, Phila., Pa. Newbrun, E., Polyusaccharide Synthesis in Plaque;Microbiol Aspects of Dental Caries, Vol III, (Supp. MicrobiologyAbstr.), 1976, pp 649-664). These compounds include the presence ofsulphur and become unstable in the presence of high oxygen compounds.The oxygen splits the sulphide bonds to form sulphates or SO₂.

Clinical observations by the inventor have led to the conclusion thatall of these biochemical compounds are attacked to a greater or lesserextent by stabilized chlorine dioxide. Since these compounds may be usedas nutrients for bacteria, the reduction of the compounds will inhibitbacterial growth. More specifically, the stabilized chlorine dioxideoxidizes carbohydrates, chondroitin sulphates, glucosaminglycans,glycoproteins, proteins and lipids. Since these compounds arise asbacterial by products and debris from dead and dying cells, are ofsalivary origin and are the mechanism of agglutination of the plaquemass, their degradation/oxidation retards plaque growth.

The initial bacterial residents of the plaque mass are aerobic, oxygenusing organisms. The saliva bathing the plaque matrixes the source ofoxygen. As the plaque thickens, the deeper layers have a reduced oxygencontent. The thicker the aerobic population of plaque matrix, the lowerthe oxygen level in the saliva. This permits the deeper layers of theplaque matrix to develop an anaerobic population of bacteria (Globerman,D. Y., and Kleinberg, I., Intra-Oral PO₂ and It's Relation to BacterialAccumulation on the Oral Tissues; Saliva and Dental Caries, (Sp. Supp.Microbiology Abstr.) 1976 pp. 275-292).

Clinical observations by the inventor lead to the discovery that the useof stabilized chlorine dioxide as a mouth rinse will rise the level ofoxygen in the saliva. The raised level of oxygen within the plaquematrix will inhibit anaerobic bacterial growth. As periodontitis iscaused by anaerobic bacteria, the potential for the development ofperiodontitis is reduced by stabilized chlorine dioxide as a rinse.

The inhibition of acquired pellicle formation, the prevention ofbacterial agglutinization and the oxidization of the plaque mass throughrinsing with chlorine dioxide in aqueous solution are independent of thegermicidal capacity of such solution. Furthermore, these factors incombination with the bacteriocidal capacity of chlorine dioxide inaqueous solution renders the solution an effective pellicle and plaqueinhibitor.

The permeability of sublingual mucous, tissue within the mouth isincreased substantially by exposure to hydrogen sulfide (H₂ S) andmethyl mercaptan (CH₃ SH). (Gaffer and Rizzo papers referenced in"Effect of Hydrogen Sulfide and Methyl Mercaptan on the Permeability ofOral Mucosa, J. Dent. Res. 63(7), July, 1984, pages 994-997).Accordingly, the toxic bacterial products attendant plaque which producethese compounds have a related effect on tissue permeability. Sincechlorine dioxide breaks the di-sulphide bonds of both these compounds,the use of chlorine dioxide in aqueous solution as a mouthwash wouldreduce the penetration potential of pathogenic materials. Evidenceexists that endotoxin and lipopolysaccharide from gram negative bacteriaare the worst of the products to penetrate the tissues. Application ofendotoxin to gingiva has caused gingival inflammation. (ibid).

Chlorine dioxide in aqueous solution used in treatment of plaque actsupon attendant gram negative bacteria. Thereby, the inventor has learnedthrough experimentation and observation that Chlorine dioxide can be apreventative product leading to oral health.

EXAMPLE I Deodorizing Mouthwash

In an effort to find a suitable control agent for mouth odor, attentionwas directed towards the use of chlorine dioxide. The characteristics ofstabilized chlorine dioxide which make it especially useful is that itis antiseptic, a bactericide, generally colorless, odorless, highlystable and has not apparent detrimental or deleterious effect on humansat the concentrations involved. As pointed out above, mouth malodor isprimarily caused by volatile sulfur compounds, such as hydrogen sulfide,methylmercaptan and dimethyl mercaptan. As pointed out above, thesechemicals are produced as degradation products of microorganisms actingon exogenous and endogenous proteinaceous substrates, oral epithelium,food debris and saliva. In order to control mouth odor, a deodorizingmouth wash consisting of a solution of 0.02% chlorine dioxide indeionized water was utilized as a rinse. Evidence indicates efficacy atlesser dilutions to 0.005% with more rapid effect at dilutions to 0.2%.Sulfides are readily oxidized by chlorine dioxide. Bacteria implicatedin the production of malodor were also effectively controlled.Inhibition of these microorganisms will reduce dental plaque formationand maintenance process.

The chlorine dioxide mouthwash or rinse solution serves to attackproduction and origin of malodor from the mouth by splitting the sulfidebonds of both hydrogen sulfide and methylmercaptan. Therefore, deliveryof stabilized chlorine dioxide provides reduction and elimination ofthese odors. Further, the bacteriostatic, bactericidal, fungistatic andfungicidal activity of stabilized chlorine dioxide will reduce thenumber of microorganisms which assist in the production of oral debrisleading to disintegration of the organic compounds ultimately producinghydrogen sulfide and methylmercaptan. The known organisms includestaphylococci, B. subtilis, B. byrocaneous, Colon bacilli, BlackPigmented Bacteriodes, Clostridia, B. sporogenes, B. histolyticum, andT. mucosum.

The mouthwash may be delivered as a simple rinse which bathes thetongue. Literature indicates that over 50% of mouth odor originates onthe mouth and tongue surface, particularly the posterior dorsal surfaceof the tongue. Accordingly, a rinse is an effective treatment. However,persons with periodontal involvement may have an increase in oralmalodor from disintegrated epithelial cells. A mouth rinse will notpenetrate to attack gingival crevicular odorizers. To optimize treatmentwith a mouthwash containing stabilized chlorine dioxide, the wash mustbe delivered into the periodontal pockets as well as dorsal and lingualsurfaces of the tongue. The preferred treatment to accomplish this isachieved by inserting the delivery tip of a syringe into the pockets orgingival crevices or by administering the wash by a mechanically poweredwater irrigating device such as one of the type sold under the trademark"Water Pik", manufactured by Teledyne Corp. Following irrigation, theuser can swish the wash throughout the mouth, covering the dorsalsurface of the tongue and other areas.

To improve the taste and appearance of the chlorine dioxide solution,appropriate sweeteners and colorings such as saccharin, peppermint andFTC #3 coloring agent may be added as is common with commerciallyavailable mouthwashes and is well-known to those in the art.

EVALUATION OF MOUTHWASH CONTAINING CHLORINE DIOXIDE FOR ITS EFFECT ONVOLATILE SULFUR COMPOUNDS

The test mouthwash which had a concentration of 0.05%, was dispersed in3/8 oz. aliquots in individual plastic containers. The study wasperformed over a 3-hour period on six human subjects with objectionableearly morning concentrations of volatile sulfur compounds (VSC) greaterthan 0.5 ng CH₃ SH/10 ml mouth air.

Rinsing Procedure: Following initial early morning VSC analysis on theday of evaluation, subjects were instructed to rinse, with vigorousswishing of rinse between teeth, for 30 seconds with 3/8 oz volumes ofthe test mouthwash. After the rinse was expectorated, the mouth wasrinsed for 30 seconds with 15 ml of 18 megavolt pure water.

VSC Analysis: All VSC analysis were performed in duplicate on eachsubject at the following times:

1. Initial screening to select subjects with objectionable early morningconcentrations of VSC.

2. On the day of evaluation, analysis were performed on early morningmouth air samples before rinsing. These values served as controls. Thus,each subject served as his own control against which the effect of therinse was calculated. Immediately following these analysis, the subjectsrinsed and were re-analyzed, 3 min., 13 min., one hour, two hours and 3hours post-rinsing. The results are summarized on the following table:

                                      TABLE 8                                     __________________________________________________________________________    SUMMARY MOUTH AIR VSC REDUCTION                                                             One Hour       Two Hour        Three Hour                       Early Morning    %       %      %       %       %         %                   Object                                                                              H.sub.2 S*                                                                       CH.sub.3 SH*                                                                       H.sub.2 S*                                                                       Red.                                                                             CH.sub.3 SH*                                                                       Red.                                                                              H.sub.2 S*                                                                       Red.                                                                             CH.sub.3 SH*                                                                       Red. H.sub.2 S*                                                                       Red. CH.sub.3 SH*                                                                       Red.                __________________________________________________________________________    1     0.82                                                                             0.72 0.39                                                                             52.44                                                                            0.37 48.61                                                                             0.39                                                                             52.44                                                                            0.59 18.06                                                                              0.75                                                                             8.54 1.08 +50                 2     1.30                                                                             1.04 0.29                                                                             77.69                                                                            0.17 83.65                                                                             0.41                                                                             68.46                                                                            0.24 76.92                                                                              0.45                                                                             65.38                                                                              0.23 72.88               3     0.98                                                                             0.77 0.77                                                                             21.43                                                                            0.57 25.97                                                                             0.69                                                                             29.59                                                                            0.52 32.47                                                                              0.82                                                                             16.33                                                                              0.62 19.48               4     0.73                                                                             1.04 0.39                                                                             46.58                                                                            0.2  75.00                                                                             0.37                                                                             49.32                                                                            0.31 70.19                                                                              0.47                                                                             35.62                                                                              0.46 55.76               5     1.56                                                                             0.88 0.58                                                                             62.82                                                                            0.41 53.41                                                                             0.75                                                                             51.92                                                                            0.40 54.55                                                                              0.86                                                                             44.87                                                                              0.80 9.09                6     1.12                                                                             1.41 0.40                                                                             64.29                                                                            0    100 0.41                                                                             63.39                                                                            0    100  0.55                                                                             50.89                                                                              0.43 69.50               Average                                                                       Reduction        54.21   64.44  52.52   58.70   36.94     30.25               __________________________________________________________________________     mg/10 ml volume of mouth air                                             

The effectiveness of chlorine dioxide was tested both in vivo and invitro and demonstrated that stabilized chlorine dioxide will kill at the99% level in ten seconds Streptococcus mutans, the principle organismimplicated in the etiology of dental caries as well as other strains oforganisms as demonstrated by the following tests:

    ______________________________________                                        IN VITRO                                                                      THE BACTERICIDAL EFFECT OF CLO.sub.2                                          AGAINST STREPTOCOCCUS MUTANS                                                  200 ppm ClO.sub.2         SURVIVED                                            pH of  Treatment   Organisms/ No.                                             Medium Seconds     0.2 ml     Organisms                                                                             % Kill                                  ______________________________________                                        4.80   5           40,000     68      99.83                                          10          40,000     116     99.96                                          20          40,000     5       99.99                                   5.95   5           40,000     1336    99.66                                          10          40,000     98      99.76                                          20          40,000     101     99.75                                   5.01   5           29,600     *TNTC   0.0                                            10          29,600     125     99.58                                          20          29,600     70      99.76                                   6.06   5           29,600     *TNTC   0.0                                            10          29,600     *TNTC   0.0                                            20          29,600     122     99.59                                   5.06   5            9,400     744     92.1                                           10           9,400     176     98.1                                           20           9,400     44      99.5                                    6.02   5            9,400     1248    86.7                                           10           9,400     920     90.2                                           20           9,400     640     93.2                                    ______________________________________                                         *Too numerous to count                                                   

MATERIALS AND METHODS

Materials used in all experiments:

1.0 AC 5215 Odorid, ClO₂ 1000 ppm, Biocide Chemical Co.

Norman Ok

1.1 Chlorine-free distilled water employed throughout

1.2 Stirring apparatus, magnetic mixer with magnetic bar; IEC

Centrifuge 6000

1.3 Petri plates (12×50 mm, 15×100 mm)

1.4 HCl 0.1 N. NaOH 0.1 N

1.5 Sodium thiosulfate solution 15%, employed 0.04 ml

1.6 Orthotolidine (o-toluidine) JT Baker, Baker Grde, boiling point200°-201° C.

C₄, C₆, H₄, HN₂, Standard Methods for Examination of

Water and Wastewater, 14th Ed. 1975 Neutral

Orthotolidine Regent, 0.04 ml employed

1.7 Diluent, saline with 0.5% Tween 80

Materials used in individual experiments:

1.0 Exp. Streptococcus mutans ATCC #27152

1.1 Brain Heart Infusion Broth employed for initial culture

1.2 Plate counts performed on plate count agar.

2.0 Exp. Bacteroides gingivalis ATCC #33277

2.1 Anaerobic Tryptic Soy Agar (TSA) with 5% sheep blood employed forinitial isolation.

2.2 Plate counts were performed on anaerobic TSA with 5% horse serum

3.0 Exp. Bacteroides melaninogenicus ATCC #15930

3.1 Anaerobic TSA with 5% sheep blood was employed throughout

3.2 Extended time interval for stirring of organisms was 30 seconds

4.0 Exp. Actinobacillus actinomycetuemocomitans ATCC #29522

4.1 Initial cultures prepared on chocolate agar

4.2 Plate counts were performed on anaerobic TSA without sheep blood

Methods:

Initially each ATCC culture employed was grown on the media documentedunder each organism. After isolation, all cultures were maintained onappropriate media. The initial bacterial count was determined by platingten-fold serial dilutions of the selected organism in its respectivemedium. After incubation, the bacterial colonies were counted and 0.2 mlof the selected dilution was employed against ClO₂. ClO₂ was employed at200 ppm. 0.8 ml of ClO₂ was mixed with 0.2 ml of organisms suspensionand mao=mixed for the selected length of time in seconds: 5, 10, 20 and30. Two organism--ClO₂ mixtures were mixed by a 45° tilting rotation ina small tube for the selected period of time.

In each experiment, subsequent to each mixing time of ClO₂organism-mixtures, excess ClO₂ was neutralized by the addition of 0.04ml of sodium thiosulfate. To assure complete neutralization of excessClO₂ has occurred, 0.04 orthotolidine was added to each ClO₂--organism-sodium thiosulfate mixture. When ClO₂ is neutralized, themixture remains clear. If residual ClO₂ is present, the mixture turnyellow after the addition of orthotolidine. Additional controls todetermine the effect of each reagent singly or in combination againsteach organism include solidum thiosulfate-organism mixtures and sodiumthiosulfate orthotolidine organism mixtures. A control plate countwithout reagents was included for each organism.

All cultures except Streptococcus mutans were grown anaerobically in COat 37° C. for 48-96 hours. Streptococcus mutans were grown aerobicallyat 37° C. for 48 hours.

IN VIVO CHLORINE DIOXIDE EVALUATION

Thirty-nine periodontal pockets in twenty-nine patients were examined bydark field and phase microscopy. The motility and density of bacteriawere evaluated from zero to three with zero being no activity and threevery active.

Of the thirty-nine teeth thirty were molars, three were bicuspids andsix were in the anterior region. Pocket dept ranged from 4 to 12millimeters.

The patients were instructed to use a 0.1% chlorine dioxide solutiontwice daily. Four of the patients used chlorine dioxide as a mouth rinseand twenty-five used it as an irrigant with monoject 412 twelve ccsyringe.

The findings follow:

    __________________________________________________________________________    CLINICAL EFFECT OF .1% CHLORINE DIOXIDE                                                                          BEFORE                                                                              AFTER                                                 BEFORE                                                                              AFTER       DARK  DARK                                 TOOTH #                                                                             CODE #                                                                             SURFACE                                                                             PHASE PHASE                                                                              % CHANGE                                                                             FIELD FIELD                                                                              % CHANGE                        __________________________________________________________________________    14    001  M     2     0    100%   3     1+   50%                             23    001  D     +     0    100%   2     +    75%                             30    002  D     0     0    0%     1     0    100%                            18    003  D     2     0    100%   2+    1-   70%                             15    004  L     +     0    100%   +     +    0%                              18    004  D     2     0    100%   2+    1    60%                             14    005  M     2     1    50%    2     2    0%                              30    005  L     1     0    100%   2     0    100%                            15    006  L     0     0    0%     2     0    100%                            19    007  D     0     0    0%     3     0    100%                            31    008  B     2     0    100%   2+    0    100%                            7     009  D     3     0    100%   3     2    33%                             2     010  M     1     0    100%   2     0    100%                            4     011  M     0     0    0%     2     0    100%                            15    011  D     1     0    100%   3     1-   75%                             3     012  M     2     1-   63%    3     2    33%                             14    012  M     2     1    50%    3     2    33%                             18    013  M     0     0    0%     3     1    67%                             3     014  M     0     0    0%     1     0    100%                            2     015  M     2     1    50%    3     2    33%                             2     015  D     2     +    75%    3     +    83%                             __________________________________________________________________________

    __________________________________________________________________________    CLINICAL EFFECT OF .1% CHLORINE DIOXIDE                                                                          BEFORE                                                                              AFTER                                                 BEFORE                                                                              AFTER       DARK  DARK                                 TOOTH #                                                                             CODE #                                                                             SURFACE                                                                             PHASE PHASE                                                                              % CHANGE                                                                             FIELD FIELD                                                                              % CHANGE                        __________________________________________________________________________    80    016  D     0     0    0%     2+    0    100%                            14    017  M     1     0    100%   3     2    33%                             3     018  M     1     0    100%   1     0    100%                            32    019  D     1+    0    100%   2     0    100%                            31    020  B     2     +    75%    3     +    83%                             2     021  M     2+    1    60%    3     2    33%                             32    022  D     1     0    100%   1     +    50%                             31    023  M     1     0    100%   3     0    100%                            3     024  D     2     0    100%   2     0    100%                            15    025  D     2     0    100%   3     1    67%                             26    025  D     0     0    0%     3     1    67%                             4     026  M     2     0    100%   2+    1-   70%                             12    026  M     1     +    50%    3     +    83%                             8     027  B     1     0    100%   2     1    50%                             3     028  M     1     0    100%   3     1    67%                             31    029  M     1     +    50%    3     1+   50%                             11    030  D     +     0    100%   1     +    50%                             __________________________________________________________________________

    ______________________________________                                        EVALUATION DATA                                                               ______________________________________                                        Phase                                                                         (Thirty Pockets with Activity)                                                Number of Pockets                                                                             % Resolution                                                                             % of Total                                         ______________________________________                                        21              100%       70%                                                2               75%        6.67%                                              1               63%        3.33%                                              1               60%        3.33%                                              5               50%        16.07%                                             ______________________________________                                        Dark Field                                                                    (Thirty-nine pockets with Activity)                                           Number of Pockets                                                                             % Resolution                                                                             % of Total                                         ______________________________________                                        14              100%       35.89%                                             3               83%        7.69%                                              2               75%        5.13%                                              2               70%        5.13%                                              4               67%        10.26%                                             1               60%        2.56%                                              5               50%        12.82%                                             6               33%        15.38%                                             ______________________________________                                         Mean resolution                                                               (All Bacterial activity stopped or was reduced).                              (Two of the pockets exhibited no reduction in bacteria after the use of       Chlorine Dioxide)                                                        

EXAMPLE II Toothpaste

As demonstrated above, stabilized chlorine dioxide can be an effectiveagent on odor producing microorganisms and enzymes. However, theeffectiveness of chlorine dioxide can be enhanced when included as aningredient of a toothpaste. Toothpaste is more effective than a rinse ofremoving malodor from the gums or gingiva. The action of the brushdislodges dead cells and putrescent debris from the gingival crevices aswell as on the various mouth surfaces and on the tongue. The chlorinedioxide contained in the toothpaste acts as discussed above to preventmalodor and serve as a deodorizer by attacking hydrogen sulfide andmethylmercaptan. A typical toothpaste would have the followingcomposition: stabilized chlorine dioxide approximately 0.005% to 0.2%;detergent polishing agent; calcium carbonate or silica gel; flavoring;saccharin; peppermint; coloring agent.

These other ingredients may vary and are the basic ingredients in manytoothpastes as is well-known to those in the art. Other formulationsincluding chlorine dioxide as the active ingredient would work as well.

EXAMPLE III Anti-Plaque Agent

Dental plaque, as mentioned above, is formed by a combination of actionsbeginning with acquired pellicle from saliva coating the tooth and asubsequent adhesion to the coating by streptococcus organisms. S. mutansdegrade sucrose into glucose or fructose which are then compounded intodextrans and levans. The dextrans act as a nutrient substrate for thegrowth of additional organisms and the production of acids whichdemineralize tooth enamel and dentin causing tooth decay. The stabilizedchlorine dioxide reacts with sulphated glycoprotiens to inhibit orreduce pellicle formation through oxidation of the sulphide bonds.Streptococcus sanguis more frequently than streptococcus mutans willadhere to pellicle to provide dextrans by way of glucocyltransferases.Chlorine dioxide is lethal to streptococcus mutans in vitro andmaterially reduces their numbers in vivo. The reduction of motility andmytosis by chlorine dioxide will reduce the amount of plaque formation.Dental plaque formation subsequent to any acquired pellicle is reducedwhen the microbial content of the mouth is reduced. Thus chlorinedioxide is an effective anti-microbial agent which functions as a dentalplaque inhibitor or retardant and as an anti-cariogenic agent. Preferredconcentrations are in the range from 0.005% to 0.2% in aqueous solutionsas for example, in deionized water with suitable coloring and flavoringsfor patient comfort.

EXAMPLE IV Anti-Gingivitis, Anti-Periodontitis and Gingival BleedingPreventative

Gingivitis and the various forms of periodontitis are known to be causedby bacteria. Principal forms implicated are Black Pigmented Bacteroidesand Actinobacillus actinomycetumcomitans. Gingivitis occurs from toxinsproduced by the aerobic bacteria in coronal dental plaque andperiodontitis occurs from toxins produced by anaerobic bacteria ininfection extending into periodontal pockets or spaced between thegingiva (gums) and the tooth root. Thus, control of gingivitis is byoxidation of compounds produced as bacterial by products that otherwisewould be the mechanism of agglutination of the plaque mass in thecoronal dental plaque and control of periodontitis by raising the levelof oxygen in the saliva to raise the level of oxygen in the plaquematrix and inhibit anaerobic bacterial growth in the plaque matrix foundin the gingival crevices and elsewhere.

By the use of an oxidizing agent of the strength of chlorine dioxide inthe mouth it will increase the amount of aerobic bacterial populationwhich will prevent the accumulations of anaerobic populations in thehigher oxygen tension. Since the anaerobes stimulate the immunereaction, which include lymphocyte cloning and subsequent release of thecompliment cascade to induce inflammation and bone loss, the chlorinedioxide will help prevent periodontitis through it's higher oxygentension created in the saliva. Further in this particular application,the higher oxygen tension will provide more oxygen available to convertthe adjacent tissues from anaerobic into aerobic glycolysis. This willincrease the number of adenosinetriphosphate molecules to increase theenergy available for adjacent cells. The covering epithelial cells arethe source of intracellular cementing glycoproteins and proteoglycans.If the cells do not have enough oxygen to function in aerobicglycolysis, there will be 1/18 the ATP production and with it theinterference of active transport (delivery of nutrition to the cells) aswell as cell adhesion to hold an effective covering and minimizebacterial penetration into the underlying connective tissues.

Clinical evidence had documented improvement in treatment of the abovediseases when stabilized chlorine dioxide is used. The organismscurrently implicated in the above are listed as follows:

1. Gingivitis, Actinomyces forms including Actinomyces, Israeli, Coccusforms,

2. Acute Necrotizing Ulcerative Gingivitis, Spirochetes, Bacteroidesintermedius, Fusiform nucleatum,

3. Juvenile Periodontitis, Actinobacillus actinomycetumcomitans,Capnocytophagia, Bacteroides intermedius,

4. Adult periodontitis, Bacteroides gingivalis, Bacteroides intermedius,Actinobacillus actinomycetumcomitans, Vibro nucleatum, Fusobactiumnucleatum, Fusobactium bacteroides, Anaerobic cocci.

Research has demonstrated that stabilized chlorine dioxide is lethal toBacteriodes gingivalis and Actinobacillus actinomycetumcomitans in vitroat the 95% level in twenty seconds with a 0.02% concentration. Researchin vivo demonstrates that these organisms are significantly reduced oreliminated in humans when chlorine dioxide agent is applied to thepocket area using a syringe or water injection device with a needle toforce penetration into the gingival crevices with the chlorine dioxideconcentration in the range of 0.05% to 0.2%. Both gingivitis andperiodontitis cause an increase in the rate of epithelial cellssloughing, aggravate oral malodor and cause some ulceration of tissueleaving the gingival bleeding; such bleeding is also reduced bytreatment with a solution of stabilized chlorine dioxide throughsplitting of the di-sulphide bonds of hydrogen sulphide and methylmercaptan.

Stabilized chlorine dioxide in aqueous solution is thus highly useful inthe treatment of gingivitis, periodontitis and bleeding gingiva.

EXAMPLE V Denture Soak

The malodors of the mouth result substantially from the volatile sulfurcompounds which are present in saliva. Saliva coats and penetratesdental prosthetic devices including full dentures and partial denturesand forms an acquired pellicle. Further, food and other cellular debrisadheres to dental prosthesis. Both anaerobic and aerobic bacteriaaccumulates on and in the microscopic faults and pores of theseprosthetic devices for form a plaque matrix, as discussed above.Stabilized chlorine dioxide in aqueous solution has been demonstrated asa bactericide. It is also effective for neutralizing sulfur-basedmalodors, removing organic debris from dental prosthesis and as adisinfectant. As a dental soak the solution is antimicrobial, removessulfur conpounds and breaks down organic material and can be used insolution form having a concentration of from approximately 0.002% to0.27%.

EXAMPLE VI Cellular Debridement Agent

Many wounds and desquamative diseases such as Lichen Planus,Desquamative Gingivitis and desquamative dermatological disease areaided by organic debridement agents and antimicrobial agents. Solutionor composition containing stabilized chlorine dioxide in aqueoussolution in 0.05% to 0.1% and higher concentrations is effective totreat these problems. One particular application would be inveterinarian applications for the purpose of reducing odor attendant tothese wounds and diseases.

EXAMPLE VII Sanitizer and Cold Sterilization Agent

The known bacterial, fungicidal and viralcidal characteristics ofchlorine dioxide also make it extremely useful as a sanitizer which canbe a solution in which materials can be dipped or by application in anaerosol spray. The sanitizer can be used for food, sickroom use bathroomand cold sterilization of many instruments and pieces of equipment notgenerally amenable to autoclave sterilization. Again, the concentrationof the stabilized chlorine dioxide would be preferably in the range offrom 0.005% to 2.0%.

EXAMPLE VIII Contact Lens Soak

Contact lenses accumulate bacteria and cellular debris from the eye. Theknown bactericidal, fungicical and viralcidal capacity of stabilizedchlorine dioxide along with its low toxicity makes stabilized chlorinedioxide solution an ideal lens soak. In addition, the capacity todegrade organic debris helps keep the lens clean and nonirritating. Thepreferred range of concentration is 0.005% to 0.2% in sterilized water.

It will be seen from the foregoing that stabilized chlorine dioxide insolution or as part of a composition or compound is effective intreating and preventing the formation of mouth malodor, inhibitingacquired pellicle and as a suitable plaque control agent, a bactericide,viralcide and fungicide superior to other compositions used today.Stabilized chlorine dioxide has been used for many years in other areasand extensive study in animals and in man have demonstrated its lowtoxicity and safety. Chlorine dioxide is approved by the EnvironmentalProtection Agency for water purification, food preparation andpreservation as well as a bacteriostatic, fungistatic and viralstaticagent.

I claim:
 1. A method for preventing periodontitis in the oral cavity,said method comprising the step of decreasing the anaerobic bacterialpopulation and increasing the aerobic bacterial population within theoral cavity by applying stabilized chlorine dioxide in aqueous solutionhaving a concentration of at least 0.005%.
 2. The method as set forth inclaim 1 wherein said step of decreasing includes the step of creating ahigher oxygen tension in the saliva present in the oral cavity.
 3. Themethod as set forth in claim 2 including the step of converting livingtissues adjacent oxygenated saliva from anaerobic into aerobicglycolysis.
 4. The method as set forth in claim 3 including the step ofincreasing the number of adenosinetrephosphate molecules.
 5. The methodas set forth in claim 1 including the step of retarding the immunereaction within the oral cavity and concomitant lymphocyte cloning. 6.The method as set forth in claim 5 including the step hindering thecompliment cascade normally resulting from lymphocyte cloning to reduceinflammation and bone loss within the oral cavity by.
 7. The method asset forth in claim 1 including the step hindering the compliment cascadenormally resulting from lymphocyte cloning to reduce inflammation andbone loss within the oral cavity by.
 8. The method as set forth in claim1 wherein the concentration of stabilized chlorine dioxide is in therange of 0.005% to 0.2%.